a Plant expression vector pUb-DR-T, pUb-D-T, and pUb-R-T. pUb1 ubiquitin promoter, SP signal peptide, tNos 3′ untranslated terminator region of the Agrobacterium tumefaciens nopaline synthase gene, p35S CaMV35S promoter, hpt hygromycin phosphotransferase. b Total genomic DNA extracted from transgenic plants was amplified by PCR using specific primers for polyprotein (521-bp) coding region. Lane 1 negative control (untransformed Pusa basmati 1); lane 2 positive control (pUbI-DR-T plasmid); lane 3, 5, and 6 plants transformed with polyprotein gene construct; lane 4 1 kb ladder. c Southern blot analysis of KpnI-digested genomic DNA from untransformed and representative polyprotein-transformed transgenic plants in T2 generation. Genomic DNA (10 µg) was digested with restriction enzymes and blotted onto nitrocellulose membranes after electrophoresis in a 0.8% agarose gel. The membrane was hybridized with 521-bp polyprotein coding sequence; lane 1 1 kb Ladder; lane 2 DR-T-18; lane 3 DR-T-2; lane 4 DR-T-14; lane 5 DR-T-11; lane 6 untransformed negative control; lane 7 DR-T-6; lane 8 DR-T-17; lane 9 DR-T-4; lane 10 DR-T-22.